Amino Acid Levels Research Articles

In vitro antioxidants and anti-inflammatory potentials of high protein-fibre cookies produced from whole wheat, sweet potato, rice bran and peanut composite flour blends

The growing demand for low-cost and functional snacks in many developing nations called for interest in the use of locally grown crops as substitutes for costly imported wheat flour. The amino acid composition, antioxidant and anti-inflammatory properties of the cookies from whole wheat, sweet potato, rice bran and peanut (56.25:18.75:5:20; 37.50:37.50:5:20; 18.75:56.25:5:20% as WPRG 1, WPRG 2, WPRG 3) composite flour blends, respectively, were obtained in this study. The 100% whole wheat and 100% refined flours served as control 1 and 2, respectively. The level of hydrophobic and aromatic amino acids was significantly (p < 0.05) high in WPRG 2 (~ 30 and ~ 10%), respectively when compared to others. However, the branched chain amino acids and Fischer ratio was significantly (p < 0.05) high in WPRG 1 (11.40% and 1.29), respectively, which could have contributed to their improved bioactivities. Notably, the composite cookie samples WPRG 1, 2 and 3 had higher hydroxyl (73.86 − 84.16%), DPPH (76.52 − 84.60%) radical scavenging as well as ferric reducing antioxidant (0.64–0.87 mmolFe2+/mg) properties than the control samples WWF and CWF, respectively. On the contrary, the metal chelating activities of the cookies WPRG 1–3 were not significantly (p > 0.05) different from control samples WWF and CWF. The improved amino acid profile and enhanced antioxidant properties of the composite cookies might have effectively influenced their anti-inflammatory properties (IC50; < 26 μg/ml) when compared to the control samples (IC50; ~ 40 μg/ml), respectively. Hence, the cookies that comprised of antioxidants and anti-inflammatory potentials needed in human health, were acceptable by the consumers.Graphical

DAAO Mutant Sites among Different Mice Strains and Their Effects on Enzyme Activity.

Previous studies reported that D-amino acid oxidase (DAAO) activity was closely associated with neuropathic pain, cognitive characteristics of schizophrenia and so on. To determine DAAO mutant sites in different strains of mice and their effects on enzyme activity, we successfully constructed a prokaryotic expression system for heterologous expression of DAAO in vitro. There were total five nucleotide mutations distributed in exons 2, 8, 9, 10 of C57 mice. Three mutations located on exons 8 and 9 were synonymous mutations and had no variation on the encoded amino acid. The remaining two mutations in exons 2 (V64A) and 10 (R295H) were non-synonymous mutations, which might affect enzymatic activity and protein structure of mDAAO. Based on the determination of the kinetic constants and IC50 of mDAAO mutants in vitro, the differences in amino acid levels at these two sites (V64A, R295H) increased the affinity of C57 DAAO with substrate and enhanced its catalytic efficiency. Besides, the IC50 value of C57 DAAO was less than that of Balb/c and other DAAO mutants (SUN: reducted by about 11.9%; CBIO: reducted by about 26.5%), which meant that the affinity of C57 DAAO with CBIO was higher.

The Effect of Rhizobium Inoculation on the Nutritional Value of Crops in the Legume–Cereal Intercropping System in Northern Kazakhstan

In this study, the changes in yield, nutrient content, and amino acid levels in legume–cereal grass mixtures were qualitatively evaluated depending on the legume–cereal combination and inoculation with preparations based on Rhizobium. This study, taking into account the biological characteristics of legume forage crops, used inoculations with strains of nodule bacteria and associative nitrogen fixers to enhance the process of the nitrogen fixation of mixed crops of legumes and cereal. The aim of this study was to compare the yields and nutritional values of monocultures and mixed crops, as well as to determine the effects of preparations based on strains of nodule bacteria and the associated nitrogen fixer on the photosynthetic activity and yield of combined annual legume–grain crops. A comparative study of forage crop biomass was conducted to analyze crude protein, fiber, carotene, and amino acid content in monocultures and legume–cereal mixtures, with and without the use of nodule-bacteria-based preparations (Rhizotorphin, Mizorine, Flavobactrin, and Azolene). The combined effect of crop mixtures and biological products led to increased green mass yield, protein content, and feed productivity. Notably, two-component mixtures with Rhizotorphin inoculation increased green mass yield by 8.79%, while three-component mixtures saw a 16.49% increase. The oat–pea mixture showed the most significant amino acid improvements, with lysine increasing by 6.26% and tyrosine by 3.24%. The general conclusion reached by the two-year experiment of 2022–2023 in the hill–plain zone of northern Kazakhstan is that double grass mixtures treated with nodule bacteria are more productive than monoculture crops in this area. These results suggest that inoculation with bacterial strains can effectively enhance the productivity of forage crops in northern Kazakhstan, providing a basis for future recommendations on optimizing herbaceous crop combinations. It is recommended to grow annual forage crops in mixtures with legumes to produce highly nutritious feeds with high metabolic energy in terms of biochemical composition.

Analysis of the relationship between neonatal birth weight and meconium metabolites based on birth cohort metabolomics

Neonatal birth weight is a crucial indicator of intrauterine growth and development with important implications for child development and adult health. The birth weight of a newborn is closely linked to the nutrition and health of the mother during pregnancy as well as genetic factors. Therefore, assessing the metabolic status of the fetus in utero is greatly significant for understanding the mechanisms responsible for abnormal birth weight. While previous studies often analyzed the impact of maternal metabolism on fetal development using umbilical cord blood from pregnant women, such blood may not accurately reflect the actual intrauterine environment owing to the barrier function of the placenta; moreover, obtaining biological samples during the fetal period is challenging. Meconium, the first feces excreted by a newborn, provides ideal biological material for studying maternal and infant health. Metabolomics can reveal metabolic changes in living organisms by analyzing small molecules in biological samples; hence studying meconium samples using metabolomics technology is expected to reveal fetal metabolic changes during pregnancy, thereby providing new insights into fetal nutritional intake, growth, and development, as well as metabolic pathways related to birth weight. To gain a deeper understanding of the metabolic changes associated with birth weight, this study collected metabolomic data from the meconium of 484 newborns in the established Xiaogan birth cohort using an untargeted metabolomics technique based on liquid chromatography-high resolution mass spectrometry (LC-HRMS) and analyzed the association between meconium metabolites and birth weight. This cohort exhibited incidence rates of low birth weight (<2500 g) and macrosomia (>4000 g) of 3.3% and 7.2%, respectively, which were roughly equivalent to the national average. Orthogonal partial least squares discriminant analysis revealed significant differences between the meconium metabolomes of the low birth weight and macrosomic groups when compared to the normal weight group. We discovered significant distinctions between the differential metabolites of newborns of low birth weight and those of normal weight, as well as between macrosomic and normal weight newborns that point to disparate biological pathways. Newborns with low birth weight exhibited significantly lower levels of critical amino acids, such as glutamate and proline, compared to the normal weight group, which may be associated with placental dysfunction and maternal nutritional deficiencies. Conversely, the meconium of macrosomic newborns contained significantly elevated levels of hormone metabolites such as estrone that reflected the pathophysiological state associated with maternal metabolic diseases or excessive placental hormone levels. Our study suggests that the metabolomic profile of the meconium reflects the metabolic pathways and regulatory mechanisms at play during fetal growth and development, and offers potential metabolic biomarkers and directions for future in-depth research into diseases related to fetal development. However, this study was based solely on the Xiaogan birth cohort, which was limited to specific regions and populations. A multicenter, multiethnic, and multiregional study is expected to help validate the universality of our research findings.

Targeted metabolomics analysis of fatty acids in lamb meat for the authentication of paper mulberry silage as a substitute for alfalfa silage

BackgroundThe paper mulberry (Broussonetia papyrifera L.) is a valuable source of woody forage that can be used for ruminants, such as goat and lambs. However, there is limited knowledge about how paper mulberry silage affects carcass characteristics, meat physicochemical attributes, amino acid and unsaturated fatty acids in Hu lambs compared to alfalfa silage. The objective of this experiment was to investigate the impact of substituting alfalfa silage with paper mulberry silage on the slaughter performance, meat quality, free amino acid and fatty acid composition in muscles of Hu lambs.ResultsThirty 60-day-old male Hu lambs with 17.13 ± 0.26 kg body weight were randomly allocated into paper mulberry silage (30% of total mixed ration) and alfalfa silage (30% of total mixed ration) treatment, and the feeding trial lasted 70 days. The results indicated no significant differences in all measurements (P > 0.05). However, compared to the group fed with alfalfa silage, the group fed with paper mulberry silage exhibited a reduction in meat drip loss (P < 0.05) without any notable effect on meat nutrients (P > 0.05). Targeted metabolomics analysis revealed that feeding paper mulberry silage led to decreased levels of certain bitter amino acids, such as valine, leucine, isoleucine, tryptophan, and phenylalanine (P < 0.05). Furthermore, the consumption of paper mulberry silage significantly augmented the levels of monounsaturated and polyunsaturated fatty acids, particularly n6-polyunsaturated fatty acids (C18:2n6, C20:3n6, C20:4n6, etc.), in meat.ConclusionsSubstituting paper mulberry silage for alfalfa in the daily diet of Hu lambs not only has no detrimental effect on animal performance but also improves meat unsaturated fatty acid composition.Graphical

Effect of synchronicity of amino acid supply on the synthesis of protein in C2C12 myotubes cultured in vitro

Previous studies inferred that the synthesis rate/efficiency of protein in body tissue is probably affected by synchronicity of different amino acid (AA) supply in its metabolic pool. In order to further observe the influence of synchronicity of AA supply on the synthesis of protein in cell level, a cell culture experiment in vitro was conducted with C2C12 myotubes. C2C12 myotubes were cultured for 24 h, meanwhile the culture medium was replaced for each 8 h. Those myotubes were subjected to 3 treatments (1 for controlled and 2 for tested), control myotubes were cultured with same normal complete medium within the whole 24 h, and the 2 tested myotubes were cultured with asynchronous amino acid supply medium in which the levels of different AAs (Lysine, threonine, methionine, leucine, valine and glutamic acid) either increased and then decreased or decreased and then increased, at different replaced medium time point (at 0, 8, and 16 h). However, during the whole experiment period all the 3 treated myotubes received same amount of each AA. The sample of the myotubes were used for myotube morphology, protein, AA, and proteomic analysis. The results showed that asynchronous AA nutrition affect the synthesis and degradation of myotube proteins, and the AAAS in the medium increase, thus decreasing the synthesis rate of myotube proteins (p &amp;lt; 0.05) and decreasing the diameter of myotubes (p &amp;lt; 0.05). The process of reduced protein synthesis affects the PI3K-AKT and FoxO signaling pathway by downregulating the levels of IRS1 and EGFR, and the degradation amplitude is greater than the synthesis amplitude. Therefore, this study further revealed the effect of the asynchronous supply of amino acids on myotube protein synthesis and the underlying mechanism and provided a theoretical reference for the precision of nutrition to animals.

Circulating amino acid signature features urea cycle alterations associated with coronary artery disease

Coronary artery disease (CAD) remains a leading cause of death worldwide and imposes a substantial socioeconomic burden on healthcare. Improving risk stratification in clinical practice could help to combat this burden. As amino acids are biologically active metabolites whose involvement in CAD remains largely unknown, this study investigated associations between circulating amino acid levels and CAD phenotypes. A high-coverage quantitative liquid chromatography-mass spectrometry approach was applied to acquire the serum amino acids profile of age- and sex-coarsened-matched patients with CAD (n = 46, 66.9 years, 74.7% male) and healthy individuals (n = 120, 67.4 years, 74.7% male) from the COmPLETE study. Multiple linear regressions were performed to investigate associations between amino acid levels and (a) the health status (CAD vs. healthy), (b) the number of affected coronary arteries, or (c) the left ventricular ejection fraction. Regressions were adjusted for age, sex, daily physical activity, sampling, and fasting time. Urea cycle amino acids (ornithine, citrulline, homocitrulline, aspartate, and arginine) were significantly and negatively associated with CAD, the number of affected coronary arteries, and the left ventricular ejection fraction. Lysine, histidine, and the glutamine/glutamate ratio were also significantly and negatively associated with the CAD phenotypes. Overall, patients with CAD displayed lower levels of urea cycle amino acids, highlighting a potential role for urea cycle amino acid profiling in cardiovascular risk stratification.Trial registrationThe study was registered on https://www.clinicaltrials.gov (NCT03986892) on June 5, 2019.

Modified throughput ninhydrin method for the qualitative assessment of dietary protein absorption in pig plasma

Abstract Background Protein maldigestion and malabsorption lead to malnutrition and are a feature of exocrine pancreatic insufficiency (EPI). Although it is the current standard, measurement of nitrogen in stool to assess protease activity is indirect. Up to 80% of hydrolysed proteins appear in blood in the form of peptides so we developed a method to measure peptide-derived amino acids in plasma as a relevant measure of proteolysis, verified its accuracy, precision, and linearity, and validated it in a porcine model. Materials and Methods We modified a ninhydrin method. Large proteins were eliminated from plasma with 10 kDa-cut-off centrifugal filters. Free and total amino acids were measured in permeate before and after its hydrolysis. Peptide-derived amino acids were quantified by subtracting free amino acids from total amino acids. We verified the method in vitro and by comparing results in healthy and EPI pigs. Results The accuracy of the analysis was close to 100%, with excellent precision (mean relative standard deviation for low, medium and high amino acid levels = 0.88%) and with stringent linearity (r2=0.986, %RE =5.23). The high-throughput ninhydrin method detected levels of peptide-derived amino acids in vivo with maximal changes seen approximately 2 hours postprandially in young pigs. The AUC and Cmax were significantly higher in healthy compared to EPI pigs (p = 0.0026 and p = 0.0037, respectively). Conclusions The high-throughput ninhydrin method is a sensitive, reliable, and practical method for the estimation of dietary peptide-derived amino acids. This assay endpoint could serve as a direct biomarker of protein digestion and absorption.

Effects of Different LED Spectra on the Antioxidant Capacity and Nitrogen Metabolism of Chinese Cabbage (Brassica rapa L. ssp. Pekinensis)

Light quality optimization is a cost-effective method for increasing leafy vegetable quality in plant factories. Light-emitting diodes (LEDs) that enable the precise modulation of light quality were used in this study to examine the effects of red-blue (RB), red-blue-green (RBG), red-blue-purple (RBP), and red-blue-far-red (RBF) lights on the growth, antioxidant capacity, and nitrogen metabolism of Chinese cabbage leaves, while white light served as the control (CK). Results showed that the chlorophyll, carotenoid, vitamin C, amino acid, total flavonoid, and antioxidant levels of Chinese cabbage were all significantly increased under RBP combined light treatment. Meanwhile, RBG combined light treatment significantly increased the levels of amino acids but decreased the nitrite content of Chinese cabbage. In addition, RBF combined light treatment remarkably increased the amino acid levels but decreased the antioxidant capacity of Chinese cabbage. In conclusion, the addition of purple light to red-blue light was effective in improving the nutritional value and antioxidant capacity of Chinese cabbage. This light condition can be used as a model for a supplemental lighting strategy for leafy vegetables in plant factory production.

Ubiquitin regulatory X (UBX) domain-containing protein 6 is essential for autophagy induction and inflammation control in macrophages.

Ubiquitin regulatory X (UBX) domain-containing protein 6 (UBXN6) is an essential cofactor for the activity of the valosin-containing protein p97, an adenosine triphosphatase associated with diverse cellular activities. Nonetheless, its role in cells of the innate immune system remains largely unexplored. In this study, we report that UBXN6 is upregulated in humans with sepsis and may serve as a pivotal regulator of inflammatory responses via the activation of autophagy. Notably, the upregulation of UBXN6 in sepsis patients was negatively correlated with inflammatory gene profiles but positively correlated with the expression of Forkhead box O3, an autophagy-driving transcription factor. Compared with those of control mice, the macrophages of mice subjected to myeloid cell-specific UBXN6 depletion exhibited exacerbated inflammation, increased mitochondrial oxidative stress, and greater impairment of autophagy and endoplasmic reticulum-associated degradation pathways. UBXN6-deficient macrophages also exhibited immunometabolic remodeling, characterized by a shift to aerobic glycolysis and elevated levels of branched-chain amino acids. These metabolic shifts amplify mammalian target of rapamycin pathway signaling, in turn reducing the nuclear translocation of the transcription factor EB and impairing lysosomal biogenesis. Together, these data reveal that UBXN6 serves as an activator of autophagy and regulates inflammation to maintain immune system suppression during human sepsis.

Integration of metabolomics and transcriptomics reveals the toxicological mechanism of deltamethrin exposure in Chinese mitten crab Eriocheir sinensis

This study investigated the toxicological mechanism of deltamethrin on Chinese mitten crab Eriocheir sinensis juveniles in fresh water. We first conducted an acute toxicity test, followed by laboratory methods to detect changes in immune-related indices in terms of antioxidant enzyme markers, lipid metabolism-related genes, and autophagy-related and apoptosis genes. The acute toxicity (96-h LC50) of deltamethrin to E. sinensis was 7.195 μg/L. After 48 h of exposure, serum showed elevated immune-related indices (P < 0.05) for alkaline phosphatase (AKP), acid phosphatase (ACP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), complement components C3 and C4, and the key pro-inflammatory cytokines interleukin-6, interleukin-1β, and tumor necrosis factor alpha (TNF-α). In hepatopancreas at 48 h, indicators related to the antioxidant system, namely superoxide dismutase (SOD) and glutathione (GSH), were significantly elevated, whereas nitric oxide and total antioxidant capacity (T-AOC) were decreased (P < 0.05). In contrast, lipid metabolism indices for triglyceride (TG), total cholesterol (TC), and malondialdehyde (MDA) were increased (P < 0.05). Transcriptomics and metabolomics revealed that exposure to deltamethrin disrupted the lipid metabolic process in the hepatopancreas mainly by altering fatty acid synthesis, amino acid metabolism, immune signaling, and autophagy activation, while the exposure increased the content of phospholipids and cholesterol but decreased the levels of amino acids and palmitoleic acid. Quantitative genetics revealed significantly aberrantly expressed (P < 0.05) lipid metabolism-related genes, including acc1, fasn, scd1, and pnpla2, all key genes involved in lipid accumulation. Deltamethrin exposure also significantly altered (P < 0.05) gene expression levels for Toll-like receptor (tlr), myeloid differentiation factor 88 (myd88), crustin1, anti-lipopolysaccharide factor isoform 3 (alf3), tumor necrosis factor alpha (tnf-α), and NF-κB transcription factor relish. Furthermore, deltamethrin activated the toll-like receptor/major myeloid differentiation response gene 88/nuclear factor kappa-light-chain-enhancer of activated B cells (TLR/MyD88/NF-kB) signaling pathway, which activates a nonspecific immune response in E. sinensis. Additionally, carnitine palmitoyltransferase 1 A (cpt1a), cytochrome c (cyt-c), adenosine 5′-monophosphate (amp)-activated protein kinase (ampk), the autophagosomal protein microtubule-associated protein 1 light chain 3c (lc3c), and the autophagy-related proteins beclin1, atg5, atg12 were significantly induced (P < 0.05) in the adenosine monophosphate-activated protein kinase/rapamycin (AMPK/mTOR) signaling pathway. These changes resulted in excess free radicals, causing oxidative stress in the mitochondrial membrane, promoting mitochondrial autophagy. The results confirm that deltamethrin exposure can induce hepatopancreatic injury by promoting mitochondrial autophagy, activating an immune response, and inhibiting lipid metabolism. Overall, this study provides multi-level information to reveal the toxic effects of deltamethrin on E. sinensis.

Associations of Amino Acids with the Risk of Prediabetes: A Case-Control Study from Kazakhstan.

The high global prevalence of prediabetes requires its early identification. Amino acids (AAs) have emerged as potential predictors of prediabetes. This study investigates the association between amino acids and prediabetes in the Kazakh population. In this case-control study, serum AAs levels were measured using the Trace GC 1310 gas chromatography system coupled with the TSQ 8000 triple quadrupole mass spectrometer (Thermo Scientific, Austin, TX, USA) followed by silylation with the BSTFA + 1% TMCS derivatization method. Biochemical parameters, including total cholesterol, HDL-C, LDL-C, triglycerides, fasting glucose, HbA1c, and Creatinine, were assessed for each participant. Trained professionals conducted anthropometric and physical examinations (which included taking blood pressure and heart rate measurements) and family history collection. A total of 112 Kazakh individuals with prediabetes and 55 without prediabetes, aged 36-65 years, were included in the study. Only Alanine and valine showed a significant association with prediabetes risk among the 13 AAs analyzed. Our findings revealed an inverse relationship between Alanine and Valine and prediabetes in individuals of Kazakh ethnicity. A lower serum level of Alanine and Valine may serve as a predictive biomarker for prediabetes in the Kazakh population.

Is malting an absolute must? Native triticale as a stand-in for barley malt in the brewing process.

To remain competitive, brewers must innovate by incorporating novel elements beyond traditional styles. Thus, exploring triticale as a modern substitute for barley malt is promising, especially given its higher amylolytic activity compared to barley. This study aimed to assess the impact of substituting up to 50% of barley malt with unmalted triticale on green beer quality, encompassing multiple stages from wort production to primary fermentation at a laboratory scale. Triticale-based worts (ratios 10-50%) had lower extract content than 100% barley malt. However, incorporating 10% of triticale led to only a 1% decrease in extract content compared to the all-malt wort. Shearzyme® 500L, an endo-1,4-β-xylanase with β-glucanase side activity, effectively addressed wort viscosity by breaking down arabinoxylans and β-glucans in triticale cell walls. All triticale-based beers exhibited lower ethanol content compared to reference beer, as is typical when using adjuncts. In green beer, a 50% triticale ratio lowered ethanol content by 16% (without enzyme) and 19% (with enzyme) compared to 100% malt beer. However, green beer with 10% triticale had satisfactory levels of total polyphenol and vicinal diketone content, among other parameters. Commercial enzyme application significantly enhanced proteolytic activity within the grain. Fermentations of enzyme-treated worts showed higher amino acid levels, further confirming the increased proteolytic activity facilitated by the chosen enzyme. Overall, this study provides a comprehensive analysis of the brewing process using native triticale. Building on this foundation, future studies will focus on optimizing mashing conditions to enhance the fermentation profile of the wort. © 2024 Society of Chemical Industry.

Investigate the metabolic changes in intestinal diseases by employing a 1H-NMR-based metabolomics approach on Caco-2 cells treated with cedrol.

Mitochondrial dysfunction may precipitate intestinal dysfunction, while inflammatory bowel disease manifests as a chronic inflammatory ailment affecting the gastrointestinal tract. This condition disrupts the barrier function of the intestinal epithelium and alters metabolic products. Increasing mitochondrial adenosine triphosphate (ATP) synthesis in intestinal epithelial cells presents a promising avenue for colitis treatments. Nevertheless, the impact of cedrol on ATP and the intestinal barrier remains unexplored. Hence, this study is dedicated to examining the cedrol's protective effect on an inflammatory cocktail (IC)-induced intestinal epithelial barrier dysfunction in Caco-2 cells. The finding reveals that cedrol enhances ATP content and the transepithelial electrical resistance value in the intestinal epithelial barrier. Moreover, cedrol mitigates the IC-induced decrease in the messenger ribonucleic acid (mRNA)expression of tight junction proteins (ZO-1, Occludin, and Claudin-1), thereby ameliorating intestinal epithelial barrier dysfunction. Furthermore, nuclear magnetic resonance (NMR)-based metabolomic analysis indicated that IC-exposed Caco-2 cells are restored by cedrol treatments. Notably, cedrol elevates metabolites such as amino acids, thereby enhancing the intestinal barrier. In conclusion, cedrol alleviates IC-induced intestinal epithelial barrier dysfunction by promoting ATP-dependent proliferation of Caco-2 cells and bolstering amino acid levels to sustain tight junction messenger ribonucleic acid expression.

Acceptability of dried blood spot collection by primary caregivers of Filipino patients with maple syrup urine disease (MSUD) and phenylketonuria (PKU).

MSUD and PKU require lifetime management hence, regular monitoring of amino acid levels is needed to achieve good metabolic control. Ideally, plasma amino acid analysis (PLAA) is used to monitor concentrations but is expensive and not widely available in local laboratories. The newborn screening program in the Philippines uses dried blood spot (DBS) analysis as an alternative where only trained healthcare providers are allowed to perform the collection at selected facilities. With the increasing number of patients, DBS monitoring has been noted to be delayed due to multiple factors. This issue became even more evident during the COVID-19 pandemic where high-risk patients need to travel outside for blood collection. The study used a cross-sectional study design to determine the primary caregivers' perspective on DBS self-sampling for patients with MSUD and PKU and the acceptability of the samples collected. This was done through a series of collection training, pre-/post- surveys, and 10-item questionnaire, and an in-depth 1-on-1 interview for thematic analysis. The acceptability of samples was processed and evaluated by the newborn screening laboratory. At-home DBS collection by primary caregivers was found to be acceptable. The provision of knowledge and routine collection training by the medical team aids in the increase of sample acceptability as well as a source of empowerment in being equipped to take care of their child. It is highly recommended that DBS samples collected by caregivers be considered acceptable for more time and cost-saving monitoring of the patients' metabolites. This practice also promotes timely and appropriate management which can lead to better patient health outcomes.

Effect of various levels of standardized ileal digestible branched-chain amino acids on lactating sow and litter performance.

Three hundred and sixty sows were used to investigate the effect of various dietary branched-chain amino acids (BCAA) levels on sow lactation and piglet growth performance. On day 112 ± 1.4 of gestation, sows were blocked by the parity group (P1, P2, P3+) and randomly assigned to 1 of 6 dietary treatments containing various levels of standardized ileal digestible (SID) Leu, Ile, and Val. The experimental diets were formulated to the desired levels of BCAA by replacing cornstarch in a basal diet with l-leucine, l-isoleucine, and l-valine. Dietary BCAA levels relative to SID Lys were 114% or 180% for Leu, 56% or 64% for Ile, and 64% or 120% for Val. Diets were formulated to be isocaloric (3.23 Mcal ME/kg) and met or exceeded all other NRC (2012) essential amino acid and vitamin and mineral recommendations. Sow body weight (BW) and backfat thickness were measured at the time of entry into the farrowing room and at weaning. Piglet litter weights were recorded after cross-fostering and weaning to calculate the litter growth rate. Data were analyzed using generalized linear mixed models with fixed effects of dietary treatment and parity group and a random effect of lactation group. The models were fit using R (v4.4.1; R Core Team, 2024). The sow and her litter were the experimental unit, and results were considered significant if P < 0.05. On average, sows nursed their litters for 21.3 d (P = 0.998). The mean parity by treatment ranged from 3.8 to 3.9 (P = 0.999). After farrowing, the mean sow BW was 220kg with a range between treatments of 216 to 222kg (P = 0.523). On average, sows gained 2.3% of their BW (P = 0.740) with an average daily feed intake of 8.74kg/d (P = 0.903). As expected, sow Leu, Ile, and Val intakes were different across treatments (P ≤ 0.001) and corresponded to the varying dietary levels of BCAA. Sows entered farrowing with an average backfat thickness of 11.50mm (P = 0.919) and lost 6.5% backfat through lactation (P = 0.880). Sows started the trial with an average of 14.1 piglets/sow (P = 0.967) and weaned 12.7 piglets/sow (P = 0.995) with a piglet ADG of 0.22kg/d (P = 0.280) and a daily litter growth rate of 2.90kg/d (P = 0.547). In conclusion, there was no evidence of an effect of the various leucine, isoleucine, and valine levels evaluated in this study on lactating sow and piglet performance.

Proximate and Amino Acid Composition of Freshwater Fish of Nagpur, Maharashtra, India

Background: The study aimed to evaluate proximate and amino acids profile of selected freshwater species from Nagpur, India. Nutritional composition of calbasu (Labeo calbasu), silver carp (Hypophthalmichthys molitrix), tilapia (Oreochromis niloticus) and pangasius (Pangasianodon pangasius) was investigated to make aware the consumer about the present nutrient composition of these fish. Methods: Iced Calbasu, Silver carp, Tilapia and Pangasius fish were procured from Mayo fish market, Nagpur, Maharashtra, India and brought to the laboratory of Fish Processing Technology, College of Fishery Science, Nagpur. The marketable sized (average weight of 0.5-1.3 kg) fish were used for evaluation of proximate composition the methods of AOAC (2005). The amino acids profiling of fish samples were assessed by using High Performance Liquid Chromatography (HPLC, Shimadzu LC-6A). Result: The variation (p less than 0.05) in nutrient composition viz. proximate and amino acids of these fish mainly due the effect of feed composition, feeding habits, geographical condition and availability of feed etc. The major components of proximate composition, moisture (75.75-77.53%), protein (14.52-18.01%), lipids (2.45-6.71%) and ash (1.10-2.09%) were recorded. The protein content was relatively stable in all species. The carbohydrate was found to be lower than 1.5%. The energy value of the fish species was between 94.81-123.51 Kcal. The pangasius had higher energy value due to high amount of fat content. During the study, the concentrations of essential amino acids (EAA) in all fish species ranged from 25.48 g/100 g to 29.41 g/100 g, while amounts of non-essential amino acids (NEAA) ranged between 27.94 g/100 g and 31.27 g/100 g. Lysine (5.14-6.31 g/100 g) and glutamic acid (6.26-8.72 g/100 g) were the most predominant amino acids found in EAA and NEAA respectively in all species. The higher level of essential amino acids was found in tilapia fish followed by silver carp, calbasu and pangasius. The results of the present study revealed that, all species have high nutritional value in terms of proximate and amino acids to offer positive health benefits upon consumption.

Synergistic Effect of Arbuscular Mycorrhizal Fungi and Germanium on the Growth, Nutritional Quality, and Health-Promoting Activities of Spinacia oleracea L.

Arbuscular mycorrhizal fungi (AMF) and the antioxidant germanium (Ge) are promising tools for boosting bioactive compound synthesis and producing healthier foods. However, their combined effect remains unexplored. This study demonstrates the synergistic impact of AMF and Ge on the growth, metabolite accumulation, biological activities, and nutritional qualities of Spinacia oleracea L. (spinach), a globally significant leafy vegetable. Individually, Ge and AMF increased biomass by 68.1% and 22.7%, respectively, while their combined effect led to an 86.3% increase. AMF and Ge also improved proximate composition, with AMF-Ge interaction enhancing crude fiber and mineral content (p < 0.05). Interestingly, AMF enhanced photosynthesis-related parameters (e.g., total chlorophyll) in Ge treated plants, which in turn increased carbohydrate accumulation. This accumulation could provide a route for the biosynthesis of amino acids, organic acids, and fatty acids, as evidenced by increased essential amino acid and organic acid levels. Consistently, the activity of key enzymes involved in amino acids biosynthesis (e.g., glutamine synthase (GS), methionine biosynthase (MS), lysine biosynthase (LS)) showed significant increments. Furthermore, AMF improved fatty acid levels, particularly unsaturated fatty acids in Ge-treated plants compared to the control. In addition, increased phenylalanine provided a precursor for the production of antioxidants (e.g., phenols and flavonoids), through the action of the enzyme phenylalanine ammonia-lyase (PAL), resulting in improved antioxidant activity gains as indicated by FRAP, ABTS, and DPPH assays. This study is the first to show that Ge enhances the beneficial effect of AMF on spinach, improving growth and nutritional quality, with promising implications for agricultural practices.

A whole food, plant-based diet reduces amino acid levels in patients with metastatic breast cancer.

Amino acids are critical to tumor survival. Tumors can acquire amino acids from the surrounding microenvironment, including the serum. Limiting dietary amino acids is suggested to influence their serum levels. Further, a plant-based diet is reported to contain fewer amino acids than an animal-based diet. Here, we investigated the impact of a whole food, plant-based diet on lowering the serum levels of amino acids in patients with cancer. Patients with metastatic breast cancer (n=18) were enrolled in clinical trial NCT03045289 . An ad libitum whole food, plant-based diet was implemented for 8 weeks without calorie or portion restriction. We found that a whole food, plant-based diet resulted in a lower intake of calories, fat, and amino acids and higher levels of fiber. Additionally, body weight, serum insulin, and IGF-1 were reduced in participants. The diet contained lower levels of essential and non-essential amino acids. Importantly, the lowered dietary intake of amino acids translated to reduced serum levels of amino acids in participants (5/9 essential amino acids; 4/11 non-essential amino acids). These findings provide a tractable approach to limiting amino acid levels in cancer patients. This data lays a foundation for studying the relationship between amino acids in patients and tumor progression. Further, a whole-food, plant-based diet has the potential to synergize with cancer therapies that exploit metabolic vulnerabilities.

New insights into transcriptional and metabolic differences between pregnant males and embryos in seahorses

Seahorses exhibit a unique reproduction mode of male pregnancy. Successful gestation plays a crucial role in the seahorse reproduction process and the aquaculture industry. However, the substantial paternal-embryo differences in seahorses remain largely unexplored. In the present study, transcriptomic and metabolic differences between pregnant males and embryos during gestation were explored to investigate the roles of key genes and metabolites in pregnant males and embryos in seahorses. Higher levels of essential amino acids and polyunsaturated fatty acids in embryos compared to fathers may indicate their vital role in embryonic development. The high expression of immune-related genes (CCLs and CCRs) and downregulation of antioxidants in fathers may provide immune protection and reduce the immunological rejection of embryos during gestation. The transcript-metabolite correlation network during pregnancy further indicated the role of key genes and metabolites during embryonic eye (N-acetyl-L-histidine, PAX6, and FZT4) and tail (WNT5 and WNT8) development. The findings of the present study not only provide valuable data on differences between pregnant males and embryos in seahorse, but also lay foundation for future research on seahorse aquaculture.